Custom antibody development

We offer world-leading antibody development platforms based on a proprietary RabMAb® rabbit monoclonal platform, a phage display platform (AxioMx), and a next-generation sequencing platform (NGS-RabMAb®)

The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

Notes

Leptin is a secreted protein factor produced by adipocytes. It regulates energy balance and body fat deposits. Leptin deficiency in humans and mice can cause in obesity.Acute low levels of leptin triggers increased appetite and chronic leptin deficiency can block the onset of puberty.

May function as part of a signaling pathway that acts to regulate the size of the body fat depot. An increase in the level of LEP may act directly or indirectly on the CNS to inhibit food intake and/or regulate energy expenditure as part of a homeostatic mechanism to maintain constancy of the adipose mass.

Involvement in disease

Defects in LEP may be a cause of obesity (OBESITY) [MIM:601665]. It is a condition characterized by an increase of body weight beyond the limitation of skeletal and physical requirements, as the result of excessive accumulation of body fat.

Associated products

Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

Sandwich ELISA

Use at an assay dependent concentration.

Images

Example of Leptin standard curve.

Background-subtracted data values (mean +/- SD) are graphed.

Interpolated concentrations of Leptin in Human serum and plasma titrations.

The concentrations of Leptin were interpolated from the Leptin standard curve and corrected for sample dilution. The mean Leptin concentration was determined to be 6,260 pg/mL in plasma (citrate) and 1,845 pg/mL in serum.

Interpolated concentrations of Leptin in Human serum from 10 donors.

Serum from 10 apparently healthy male donors was measured in triplicate for Leptin. The mean Leptin concentration was determined to be 2174 pg/mL with a range of 443 – 6250 pg/mL (Leptin was not detected in serum from Donor 7).